Protein-encapsulated doxorubicin reduces cardiotoxicity in hiPSC-cardiomyocytes and cardiac spheroids while maintaining anticancer efficacy

Madelyn Arzt; Bowen Gao; Maedeh Mozneb; Stephany Pohlman; Romina B. Cejas; Qizhi Liu; Faqing Huang; Changjun Yu; Yi Zhang; Xuemo Fan; Amelia Jenkins; Armando E. Giuliano; Paul W. Burridge; Xiaojiang Cui; Arun Sharma

doi:10.1016/j.stemcr.2023.08.005

Protein-encapsulated doxorubicin reduces cardiotoxicity in hiPSC-cardiomyocytes and cardiac spheroids while maintaining anticancer efficacy

Madelyn Arzt, Bowen Gao, Maedeh Mozneb, Stephany Pohlman, Romina B. Cejas, Qizhi Liu, Faqing Huang, Changjun Yu, Yi Zhang, Xuemo Fan, Amelia Jenkins, Armando E. Giuliano, Paul W. Burridge, Xiaojiang Cui^*, Arun Sharma^*

^*Corresponding author for this work

Pharmacology

Research output: Contribution to journal › Article › peer-review

5 Scopus citations

Abstract

The chemotherapeutic doxorubicin (DOX) detrimentally impacts the heart during cancer treatment. This necessitates development of non-cardiotoxic delivery systems that retain DOX anticancer efficacy. We used human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs), endothelial cells (hiPSC-ECs), cardiac fibroblasts (hiPSC-CFs), multi-lineage cardiac spheroids (hiPSC-CSs), patient-specific hiPSCs, and multiple human cancer cell lines to compare the anticancer efficacy and reduced cardiotoxicity of single protein encapsulated DOX (SPEDOX-6), to standard unformulated (UF) DOX. Cell viability assays and immunostaining in human cancer cells, hiPSC-ECs, and hiPSC-CFs revealed robust uptake of SPEDOX-6 and efficacy in killing these proliferative cell types. In contrast, hiPSC-CMs and hiPSC-CSs exhibited substantially lower cytotoxicity during SPEDOX-6 treatment compared with UF DOX. SPEDOX-6-treated hiPSC-CMs and hiPSC-CSs maintained their functionality, as indicated by sarcomere contractility assessment, calcium imaging, multielectrode arrays, and RNA sequencing. This study demonstrates the potential of SPEDOX-6 to alleviate cardiotoxic side effects associated with UF DOX, while maintaining its anticancer potency.

Original language	English (US)
Pages (from-to)	1913-1924
Number of pages	12
Journal	Stem cell reports
Volume	18
Issue number	10
DOIs	https://doi.org/10.1016/j.stemcr.2023.08.005
State	Published - Oct 10 2023

Funding

Funding provided by the American Heart Association Career Development Award 856987 (to A.S.); the National Institutes of Health grants 2R01CA151610 and R21CA280458 , Department of Defense grant W81XWH-18-1-0067 , Uretsky BRCA Research Fund and Samuel Oschin Cancer Institute Research Development Fund (to X.C.); the Cedars-Sinai Cancer Center fund (to A.S and X.C.); and the Fashion Footwear Charitable Foundation of New York, Inc., the Margie and Robert E. Petersen Foundation , and Linda and Jim Lippman Fund (to A.E.G.). A.S. is supported by the Board of Governors Regenerative Medicine Institute at Cedars-Sinai, an In-Space Production Award (InSPA) from NASA , and the Donna and Jesse Garber Award for Cancer Research . A.S. and S.P. are supported by a California Institute for Regenerative Medicine (CIRM) Bridges Award. The ACTN2-GFP and Tubulin-RFP fluorescent reporter hiPSC lines were purchased from Coriell and the Allen Institute for Cell Science. The WTC-GCaMP hiPSC line was a kind gift from Dr. Bruce Conklin (UCSF) and Dr. Nathaniel Huebsch (Washington University in St. Louis). We thank the Cedars-Sinai Center for Bioinformatics and Functional Genomics core facility for their assistance with RNA-sequencing. Funding provided by the American Heart Association Career Development Award 856987 (to A.S.); the National Institutes of Health grants 2R01CA151610 and R21CA280458, Department of Defense grant W81XWH-18-1-0067, Uretsky BRCA Research Fund and Samuel Oschin Cancer Institute Research Development Fund (to X.C.); the Cedars-Sinai Cancer Center fund (to A.S and X.C.); and the Fashion Footwear Charitable Foundation of New York, Inc. the Margie and Robert E. Petersen Foundation, and Linda and Jim Lippman Fund (to A.E.G.). A.S. is supported by the Board of Governors Regenerative Medicine Institute at Cedars-Sinai, an In-Space Production Award (InSPA) from NASA, and the Donna and Jesse Garber Award for Cancer Research. A.S. and S.P. are supported by a California Institute for Regenerative Medicine (CIRM) Bridges Award. The ACTN2-GFP and Tubulin-RFP fluorescent reporter hiPSC lines were purchased from Coriell and the Allen Institute for Cell Science. The WTC-GCaMP hiPSC line was a kind gift from Dr. Bruce Conklin (UCSF) and Dr. Nathaniel Huebsch (Washington University in St. Louis). We thank the Cedars-Sinai Center for Bioinformatics and Functional Genomics core facility for their assistance with RNA-sequencing. C.Y. is a named inventor for patent applications regarding “Single Protein-Encapsulated Pharmaceutics for Enhancing Therapeutic Effects” and a shareholder of Sunstate Biosciences, LLC. The remaining authors have nothing to disclose.

Keywords

cancer
cardiomyocyte
cardiotoxicity
chemotherapy
doxorubicin
iPSC
organoids
single protein encapsulation
spheroids
stem cell

ASJC Scopus subject areas

Biochemistry
Genetics
Developmental Biology
Cell Biology

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1016/j.stemcr.2023.08.005

Cite this

Arzt, M., Gao, B., Mozneb, M., Pohlman, S., Cejas, R. B., Liu, Q., Huang, F., Yu, C., Zhang, Y., Fan, X., Jenkins, A., Giuliano, A. E., Burridge, P. W., Cui, X., & Sharma, A. (2023). Protein-encapsulated doxorubicin reduces cardiotoxicity in hiPSC-cardiomyocytes and cardiac spheroids while maintaining anticancer efficacy. Stem cell reports, 18(10), 1913-1924. https://doi.org/10.1016/j.stemcr.2023.08.005

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abstract = "The chemotherapeutic doxorubicin (DOX) detrimentally impacts the heart during cancer treatment. This necessitates development of non-cardiotoxic delivery systems that retain DOX anticancer efficacy. We used human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs), endothelial cells (hiPSC-ECs), cardiac fibroblasts (hiPSC-CFs), multi-lineage cardiac spheroids (hiPSC-CSs), patient-specific hiPSCs, and multiple human cancer cell lines to compare the anticancer efficacy and reduced cardiotoxicity of single protein encapsulated DOX (SPEDOX-6), to standard unformulated (UF) DOX. Cell viability assays and immunostaining in human cancer cells, hiPSC-ECs, and hiPSC-CFs revealed robust uptake of SPEDOX-6 and efficacy in killing these proliferative cell types. In contrast, hiPSC-CMs and hiPSC-CSs exhibited substantially lower cytotoxicity during SPEDOX-6 treatment compared with UF DOX. SPEDOX-6-treated hiPSC-CMs and hiPSC-CSs maintained their functionality, as indicated by sarcomere contractility assessment, calcium imaging, multielectrode arrays, and RNA sequencing. This study demonstrates the potential of SPEDOX-6 to alleviate cardiotoxic side effects associated with UF DOX, while maintaining its anticancer potency.",

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Arzt, M, Gao, B, Mozneb, M, Pohlman, S, Cejas, RB, Liu, Q, Huang, F, Yu, C, Zhang, Y, Fan, X, Jenkins, A, Giuliano, AE, Burridge, PW, Cui, X & Sharma, A 2023, 'Protein-encapsulated doxorubicin reduces cardiotoxicity in hiPSC-cardiomyocytes and cardiac spheroids while maintaining anticancer efficacy', Stem cell reports, vol. 18, no. 10, pp. 1913-1924. https://doi.org/10.1016/j.stemcr.2023.08.005

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T1 - Protein-encapsulated doxorubicin reduces cardiotoxicity in hiPSC-cardiomyocytes and cardiac spheroids while maintaining anticancer efficacy

AU - Arzt, Madelyn

AU - Gao, Bowen

AU - Mozneb, Maedeh

AU - Pohlman, Stephany

AU - Cejas, Romina B.

AU - Liu, Qizhi

AU - Huang, Faqing

AU - Yu, Changjun

AU - Zhang, Yi

AU - Fan, Xuemo

AU - Jenkins, Amelia

AU - Giuliano, Armando E.

AU - Burridge, Paul W.

AU - Cui, Xiaojiang

AU - Sharma, Arun

PY - 2023/10/10

Y1 - 2023/10/10

N2 - The chemotherapeutic doxorubicin (DOX) detrimentally impacts the heart during cancer treatment. This necessitates development of non-cardiotoxic delivery systems that retain DOX anticancer efficacy. We used human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs), endothelial cells (hiPSC-ECs), cardiac fibroblasts (hiPSC-CFs), multi-lineage cardiac spheroids (hiPSC-CSs), patient-specific hiPSCs, and multiple human cancer cell lines to compare the anticancer efficacy and reduced cardiotoxicity of single protein encapsulated DOX (SPEDOX-6), to standard unformulated (UF) DOX. Cell viability assays and immunostaining in human cancer cells, hiPSC-ECs, and hiPSC-CFs revealed robust uptake of SPEDOX-6 and efficacy in killing these proliferative cell types. In contrast, hiPSC-CMs and hiPSC-CSs exhibited substantially lower cytotoxicity during SPEDOX-6 treatment compared with UF DOX. SPEDOX-6-treated hiPSC-CMs and hiPSC-CSs maintained their functionality, as indicated by sarcomere contractility assessment, calcium imaging, multielectrode arrays, and RNA sequencing. This study demonstrates the potential of SPEDOX-6 to alleviate cardiotoxic side effects associated with UF DOX, while maintaining its anticancer potency.

AB - The chemotherapeutic doxorubicin (DOX) detrimentally impacts the heart during cancer treatment. This necessitates development of non-cardiotoxic delivery systems that retain DOX anticancer efficacy. We used human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs), endothelial cells (hiPSC-ECs), cardiac fibroblasts (hiPSC-CFs), multi-lineage cardiac spheroids (hiPSC-CSs), patient-specific hiPSCs, and multiple human cancer cell lines to compare the anticancer efficacy and reduced cardiotoxicity of single protein encapsulated DOX (SPEDOX-6), to standard unformulated (UF) DOX. Cell viability assays and immunostaining in human cancer cells, hiPSC-ECs, and hiPSC-CFs revealed robust uptake of SPEDOX-6 and efficacy in killing these proliferative cell types. In contrast, hiPSC-CMs and hiPSC-CSs exhibited substantially lower cytotoxicity during SPEDOX-6 treatment compared with UF DOX. SPEDOX-6-treated hiPSC-CMs and hiPSC-CSs maintained their functionality, as indicated by sarcomere contractility assessment, calcium imaging, multielectrode arrays, and RNA sequencing. This study demonstrates the potential of SPEDOX-6 to alleviate cardiotoxic side effects associated with UF DOX, while maintaining its anticancer potency.

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KW - cardiomyocyte

KW - cardiotoxicity

KW - chemotherapy

KW - doxorubicin

KW - iPSC

KW - organoids

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KW - spheroids

KW - stem cell

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Protein-encapsulated doxorubicin reduces cardiotoxicity in hiPSC-cardiomyocytes and cardiac spheroids while maintaining anticancer efficacy

Abstract

Funding

Keywords

ASJC Scopus subject areas

UN SDGs

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